SYBR GREEN 荧光定量聚合酶链反应检测粘蛋白1在乳腺癌诊断中的应用

摘要

Objective To investigate the application of SYBR GREEN fluorescent quantification polymerase chain reaction (SYBR GREEN FQ-PCR)to detect the MUC1 mRNA of circulating tumor cells in diagnosis and therapeutic monitoring of breast canc -er.Methods Withβ-actin as internal control,MUC1 mRNA in peripheral blood cell samples in 45 health women,91 patients with be-nign breast disease and 193 patients with breast cancer and 87 with other cancer patients were detected by SYBR GREEN FQ-PCR.The relationship between MUC1 mRNA and clinicopathological parameters was analyzed .Results The positive rate of MUC1 mRNA detec-tion in the patients with breast cancer was significantly higher than that in healthy individuals ,patients with benign breast disease pa-tients and other types of cancers ( P<0.05 ) .The positive rate in the patients with benign breast disease and other types of cancers was significantly higher than that in the healthy individuals ( P<0.05 ) .There was no significant difference in the positive rate between be-nign breast disease patients and other cancer patients ( P >0.05 ) .The independent factors of MUC 1 mRNA expression were clinical stage ,histological grade and axillary metastatic lymph node and HER 2eu ( P<0.05 ) .Conclusion FQ-PCR is a rapid and sensitive and specific method for quantitative detection of mucin 1 mRNA.It can be used for the diagnosis ,therapy,metastasis and prognosis of breast cancer .%目的：探讨SYBR GREEN荧光定量聚合酶链反应法检测外周血循环肿瘤细胞中粘蛋白1基因表达水平在乳腺癌诊断和治疗监测中的应用。方法以β-actin为内对照，采用SYBR GREEN荧光定量聚合酶链反应法测定45名健康女性体检者、91例良性乳腺疾病患者和193例乳腺癌患者以及87例其它肿瘤患者外周血中粘蛋白1的表达量，分析不同临床病理因素与基因表达的相关性。结果乳腺癌患者粘蛋白1阳性率显著高于健康体检者、良性乳腺疾病和其它肿瘤患者（P＜0.05）；良性乳腺疾病及其它肿瘤患者粘蛋白1阳性率显著高于健康体检者（P＜0.05），而良性乳腺疾病与其它肿瘤患者粘蛋白1阳性率差异无统计学意义（ P＞0.05）；乳腺癌临床分期、组织学分级、腋淋巴结转移及HER2／neu表达是MUC1 mRNA表达独立相关因素（ P＜0.05）。结论 SYBR GREEN 荧光定量聚合酶链反应法是灵敏较高、特异较强的快速定量检测粘蛋白1方法，可有效监测乳腺癌诊断、疗效、转移和预后。